Nol9 Is a Spatial Regulator for the Human ITS2 Pre-rRNA Endonuclease–Kinase Complex. Issue 19 (6th September 2019)

• Record Type: Journal Article
• Authors: Gordon, Jacob; Pillon, Monica C.; Stanley, Robin E.
• Is Part Of: Journal of molecular biology. Volume 431:Issue 19(2019)
• Page Start: 3771
• Page End: 3786
• Publication Date: 2019-09-06
• Subjects: NoLS nucleolar localization sequence -- PNK polynucleotide kinase -- LCT Las1L C-terminal tail -- CC coiled-coil -- HEPN higher eukaryotes and prokaryotes nucleotide-binding -- ITS2 internal transcribed spacer 2 -- pre-rRNA precursor ribosomal RNA; Las1L -- Nol9 -- endoribonuclease -- polynucleotide kinase -- nucleolar localization sequence; Molecular biology -- Periodicals; Biology -- Periodicals; Biochemistry -- Periodicals; Bacteriology -- Periodicals; Molecular Biology -- Periodicals; Biochemistry -- Periodicals; Biologie moléculaire -- Périodiques; Biologie -- Périodiques; Biochimie -- Périodiques; Moleculaire biologie; Biochemistry; Biology; Molecular biology; Periodicals; 572.805
• DOI: 10.1016/j.jmb.2019.07.007
• Languages: English
• ISSNs: 0022-2836

Abstract

Abstract: The ribosome plays a universal role in translating the cellular proteome. Defects in the ribosome assembly factor Las1L are associated with congenital lethal motor neuron disease and X-linked intellectual disability disorders, yet its role in processing precursor ribosomal RNA (pre-rRNA) is largely unclear. The Las1L endoribonuclease associates with the Nol9 polynucleotide kinase to form the internal transcribed spacer 2 (ITS2) pre-rRNA endonuclease–kinase machinery. Together, Las1L–Nol9 catalyzes RNA cleavage and phosphorylation to mark the ITS2 for degradation. While ITS2 processing is critical for the production of functional ribosomes, the regulation of mammalian Las1L–Nol9 remains obscure. Here we characterize the human Las1L–Nol9 complex and identify critical molecular features that regulate its assembly and spatial organization. We establish that Las1L and Nol9 form a higher-order complex and identify the regions responsible for orchestrating this intricate architecture. Structural analysis by high-resolution imaging defines the intricate spatial pattern of Las1L–Nol9 within the nucleolar sub-structure linked with late pre-rRNA processing events. Furthermore, we uncover a Nol9-encoded nucleolar localization sequence that is responsible for nucleolar transport of the assembled Las1L–Nol9 complex. Together, these data provide a mechanism for the assembly and nucleolar localization of the human ITS2 pre-rRNA endonuclease–kinase complex. Graphical abstract: Unlabelled Image